MCE 国际站:FITC-Dextran (MW 40000)
CAS:60842-46-8
品牌:MedChemExpress (MCE)
存储条件:4°C, protect from light *In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)
生物活性:FITC-Dextran (MW 40000) 是一种异硫氰酸荧光素 (FITC) 葡聚糖荧光探针 (Ex=495 nm; Em=525 nm)。FITC-Dextran (MW 400000) 可作为一种标记物来揭示热休克引起的细胞损伤,并研究细胞凋亡的早期和晚期阶段。FITC-Dextran (MW 400000) 还可用于细胞渗透性的研究,如血脑屏障通透性以及血脑屏障破坏程度的测定。
体外:Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs). 细胞标记[1]: 适用于正在发生凋亡的 HeLa 细胞和人外周血单个核细胞 (PBMC) (活的 HeLa 和 PBMC 不被 FITC-Dextran 染色)。 1. 43.5°C 孵育 1 h,37°C 孵育 8 h,诱导细胞凋亡。 2. 将细胞悬浮在 100 μL 的培养基中,在 Q-prep 管与 10 μL 的 propidium iodide (PI),10 μL 的 FITC-Dextran (MW 40000) 混合,PI 和 FITC-Dextran (MW 40000) 的最终浓度分别为 7.5 μM 和 1.13 μM。 3. 将细胞在室温黑暗条件下培养 25 分钟。 4. 取 3 mL 培养基的标记细胞,500 g 离心 10 分钟。 5. 取离心细胞,加入 1 mL 培养基,用流式细胞术或荧光显微镜分析 (PI: Ex=500 nm, Em=600 nm; FITC-Dextran (MW 40000): Ex=495 nm, Em=525 nm)。 细胞旁渗透性检测[4] 1. 在 transwell 室的基础培养基中加入 FITC-Dextran (0.1 mg/mL)。 2. 在 15 分钟后从 transwell 插入器中收集培养基。 3. 测量荧光信号 (Ex=485 nm, Em=538 nm)。 4. 根据荧光强度计算 FITC-Dextran 浓度。 5. 计算渗透率。 MCE has not independently confirmed the accuracy of these methods. They are for reference only.
体内:Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs). 肠屏障功能测定[5] 1. 小鼠饥饿处理 4 小时。 2. FITC-Dextran MW 40000 灌胃小鼠 (0.6 mg/g)。 3. 在 4 h 内测量荧光强度 (Ex nm/Em 520 nm)。 MCE has not independently confirmed the accuracy of these methods. They are for reference only.
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研究领域:Others
作用靶点:Fluorescent Dye
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参考文献:
[1]. Moumaris M, et al. Fluorescein isothiocyanate-dextran can track apoptosis and necrosis induced by heat shock of peripheral blood mononuclear cells and HeLa cells[J]. Open Biological Sciences Journal, 2015, 1(1). [2]. Natarajan R, et al. Fluorescein Isothiocyanate (FITC)-Dextran Extravasation as a Measure of Blood-Brain Barrier Permeability. Curr Protoc Neurosci. 2017 Apr 10;79:9.58.1-9.58.15. [3]. Eriksson I, et al. Analysis of Lysosomal pH by Flow Cytometry Using FITC-Dextran Loaded Cells. Methods Mol Biol. 2017;1594:179-189. [4]. Okabayashi K, et al. Cdc42 activates paracellular transport in polarised submandibular gland cells. Arch Oral Biol. 2021 Dec;132:105276. [5]. Yu W, et al. ACE2 contributes to the maintenance of mouse epithelial barrier function. Biochem Biophys Res Commun. 2020 Dec 17;533(4):1276-1282.